LOCUS       sequence-252688-        8625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Xylose inducible system with the CRE site for expression in B. 
            subtilis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8625)
  AUTHORS   Gonzalez LM, Mukhitov N, Voigt CA
  TITLE     Resilient living materials built by printing bacterial spores.
  JOURNAL   Nat Chem Biol. 2019 Dec 2. pii: 10.1038/s41589-019-0412-5. doi: 
            10.1038/s41589-019-0412-5.
  PUBMED    31792444
REFERENCE   2  (bases 1 to 8625)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2019 Dec 2. pii: 10.1038/s41589-019-0412-5. doi: 
            10.1038/s41589-019-0412-5."
COMMENT     Sequence Label: pLG169
FEATURES             Location/Qualifiers
     source          1..8625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1738..2454
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYKY"
     primer_bind     complement(1765..1793)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2388..2408
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3385..3403)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3471..3575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3576..3644,3645..4436)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3794..3813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4607..5195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5096..5115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatagttat tataacatgt attcacgaac gaaaatcgcc attcgccagc tgcaggaatt
       61 cgatatcaag cttctaggat ctttacattt taacgatatc tagaaaatgt tcaataacaa
      121 tggaagacat tcctaatgcc ggtgcattct ttcctaagga agatggtaat aattcatagc
      181 tattgcctaa ttggggataa accctggacg acacttcact tctaattgaa tttaaaacca
      241 taggatgtga ttcaattatg ctgtttctta aaatgatggc ttgtggattg aatgtattta
      301 gaatattagt aaggccaatt cctaaataga atccgaaatt ctgtaatgca tttaaggtgc
      361 cgatatcatt cagatgggcg aggtctatga tatcttgata ggacactttt ttctctttag
      421 tctgaagaga ttttaataaa gccttctctg aagcatacaa ttcccagcaa cctcggtttc
      481 cgcaactgca tttaggacca ttaaagtcta ttgtcatatg tcccatttct ccagagaatc
      541 cgcttacccc tctatataaa tgattgttga taataacacc gatccctata cctgtactga
      601 tactagcata aataatgtta ttgtgatttt ttgcagcacc aaatactttt tctccatacg
      661 cgccagcatt tgcctcattt tcaataaaaa caggcacatt gaacttctct tgtatgaaag
      721 attttaagtc aatatctctc cagttggagt tcggagtgaa aacaattttt tgatttttat
      781 caattagtcc aggcacgcat atgcctatac cgataagccc gtacggagat tgtggcatac
      841 gcgtaataaa gtgatgaatc atatcaatta aaatgtcttt agttatttct ggagaattcg
      901 attctaaatg gtgatgttga tcaagaatga tcgttccttc aaggtctgtt aaaatgccac
      961 taatataatc cacaccaaca tctattccta tggagtatcc tgccttttta ttaaaaacaa
     1021 gcatgacagg tcttcttccg ccactttgct gatcaagaat gatcgtccct tcaaggtctg
     1081 ttaaaatccc actaatataa tccacaccaa catctatgcc tatggagtat cctgccttct
     1141 tattaaaaac aagcatcaca ggtcttcttc cgccacttga ttgtccttga cctatttcaa
     1201 atacaagatt ttctttcatc agcgtgttta cctgtgatga aacagttgat ttatttaatc
     1261 cagtcatttc agataatttt gctcttgaaa taggtgaatt cttaagtatt tcttttaata
     1321 ataacttttg atttactttt ttgacaaagg tttgatctgc gatatccact tcatccactc
     1381 catttgtata acctttaaat taagttaaaa ttttttgtgt tcagtatgag atttagtaca
     1441 tagcgaatct tacctttatt atatctaatg tgttcatgaa aaactaaaaa aaatattgaa
     1501 aatactgatg aggttattta agattaaaat aagttagttt gtttgggcaa caaactaatg
     1561 tgcaacttac ttacaatatg acataaaatg catctgtatt tgaatttatt tttaaggagg
     1621 aaataacatg gctcaatctc attctagttc agttaactat tttgtaagcg ttaacaaagt
     1681 ggtttaatta agtcgacagc tagccgcaca attttatgta aggatggggg aattttcatg
     1741 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     1801 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
     1861 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     1921 actttcgcgt atggtcttca atgctttgcg agatacccag atcatatgaa acagcatgac
     1981 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     2041 gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga
     2101 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
     2161 tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa
     2221 gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     2281 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     2341 acacaatcta agctttcgaa agatcccaac gaaaagaggg accacatggt ccttcttgag
     2401 tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata ctagctgcat
     2461 tcagcccgcc taatgagcgg gcttttttaa ttcaatttca caatcagaga gaacgaattc
     2521 gataacccta aagttatgga aataagactt agaagcaaac ttaagagtgt gttgatagtg
     2581 cattatctta aaattttgta taataggaat tgaagttaaa ttagatgcta aaaatttgta
     2641 attaagaagg agggattcgt catgttgggg agcccgtatt tcgcgtaagg aaatccatta
     2701 tgtactattt aattctgcgt gacatcccat cgatcagacc agtttttaat ttgtgtgttt
     2761 ccatgtgtcc agtttggaat actcttaacc tcattggaaa tcgcggcata atcactggtg
     2821 gtatgattga tgaccgcgtc aacaatgacc tttatgccat attcttcagc ggctgcacac
     2881 atttctttaa attcttgttc agtacctaag taacggttgc caatttgata cgatgtcggc
     2941 tgatacagcc agtaccagtt cgacatgctt ttatctcctt gattcccttc ctttacttgg
     3001 ttaatcggag atgtctgaat ggctgtatat cctgcatcat gaatatcctt catattgtgt
     3061 tttaacgtat tgaacgacca attccatgca tgaagaatgg ttccgctttt gatcgacggt
     3121 gctgtaagct cattcgattt gttcgccgtt tcagcactcg cagccgccgg tcctgccaga
     3181 accaaatgaa acagcaataa aaatccagcg aataacggca gtaaagaggt tttgaatcgt
     3241 tttgcaaaca ttcttgacac tccttatttg attttttgaa gacttacttc ggagtcaaaa
     3301 atccctctta cttcattctt ccgcttcctc ctttcaaacc gatgtgaaga ctggagaatt
     3361 ttgttaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3421 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccat
     3541 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3601 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3661 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3721 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3781 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     3841 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3901 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3961 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4021 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4081 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     4141 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4201 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4261 ttgctgataa atctggagcc ggtgagcgtg ggtcgcgcgg tatcattgca gcactggggc
     4321 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4381 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4441 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4741 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcccgacctc
     5221 gagctggata cttcccgtcc gccaggggga catgccggcg atgctgaagg tcgcgcgcat
     5281 tcccgatgaa gaggccggtt accgcctgtt tgaggatata gtaatctttc taaatagctt
     5341 tggattggag gagtatggcc actaatacta agttcagcta ataaaaaaat ttgctaaaga
     5401 actccagctg gatttcactg atgagaatat cgtcggagat aaatataata attccacgga
     5461 ctatagacta tactagtata ctccgtctac tgtacgatac acttccgctc aggtccttgt
     5521 cctttaacga ggattgttac gtacgctaat ggcgtcaaaa caaagactct agacctaggc
     5581 cttaagatct gatcatatgc atccgcgggc ccgggttaac gcgtaatcca tggatcaaga
     5641 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggt
     5701 gccctgaatg aactgcagaa agagctggta gttggcgcac tgttcgaaga actgccgatg
     5761 tccagtaaga ttcttactat gctggttgaa ccggatgctg gtaaagctac ttgggttgct
     5821 gcttctactt atggtaccga tacaactact ggtgaggaag ttaaaggagc tcttaaagaa
     5881 atccacttca gtaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa
     5941 ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat
     6001 aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg
     6061 atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg
     6121 cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc
     6181 gcgtcgcgac taagaaaatg ccgtcaaatc cgctcgccat gacttcacta acgatgcctt
     6241 tgaaaatctt caagttcttt tctactaatt caaggcgtgt ctcaccaggt ttttggtttg
     6301 ctccggcgca aatgcagaca atatcagcat ccttgcaggg tatgtttctc tttgatgtct
     6361 ttttgtttgt gaagtatttc acatttatat tgtgcaacac ttcacaaact tttgcaagag
     6421 aaaagttttg tctgatttat gaacaaaaaa gaaaccatca ttgatggttt ctttcggtaa
     6481 gtcccgtcta gccttgccct caatggggaa gagaaccgct taagcccgag tcattatata
     6541 aaccatttag cacgtaatca aagccaggct gattctgacc gggcacttgg gcgctgccat
     6601 tattaaaaat cacttttgcg ttggttgtat ccgtgtccgc aggcagcgtc agcgtgtaaa
     6661 ttccgtctgc atttttagtc attggttttc caggccaaga tccggtcaat tcaattactc
     6721 ggctcccatc atgtttatag atataagcat ttacctggct ccaatgattc ggattttgat
     6781 agccgatggt tttggccgac gctggatctc ttttaacaaa actgtatttc tcggtcctcg
     6841 ttacaccatc actgttcgtt ccttttaaca tgatggtgta tgttttgcca aatggatctc
     6901 cttttccgat tgtgaattga tctccatcct taaacgcctg tcgtctggtc cattattgat
     6961 ttgataaacg gcttttgttg tattcgcatc tgcacgcaag gtaatcgtca gttgatcatt
     7021 gaaagaatgt gttacacctg ttttgtaatt ctcaaggaaa acatgaggcg cttttgcaat
     7081 atcatcagga taaagcacag ctacagacct ggcattgatc gtgcctgtca gtttaccatc
     7141 gttcacttga aatgaacccg ctccagcttt attgtcatac ctgccatcag gcaattttgt
     7201 tgccgtattg atagagacag aggatgaacc tgcatttgcc agcacaacgc catgtgagcc
     7261 gcgctgattc ataaatatct ggttgtttcc attcgggttc gagagttcct caggctgtcc
     7321 agccatcaca ttgtgaaatc tattgaccgc agtgatagcc tgatcttcaa ataaagcact
     7381 cccgcgatcg cctatttggc ttttccccgg gaacctcaca ccatttccgc ctccctcagg
     7441 tctggaaaag aaaagaggcg tactgcctga acgagaagct atcaccgccc agcctaaacg
     7501 gattccccct atgcaagggt ttattgtttt ctaaaatctg attaccaatt agaatgaata
     7561 tttcccaaat attaaataat aaaacaaaaa aattgaaaaa agtgtttcca ccattttttc
     7621 aattttttta taattttttt aatctgttat ttaaatagtt tatagttaaa tttacatttt
     7681 cattagtcca ttcaatattc tctccaagat aactacgaac tgctaacaaa attctctccc
     7741 tatgttctaa tggagaagat tcagccactg catttcccgc aatatctttt ggtatgattt
     7801 tacccgtgtc catagttaaa atcatacggc ataaagttaa tatagagttg gtttcatcat
     7861 cctgataatt atctattaat tcctctgacg aatccataat ggctcttctc acatcagaaa
     7921 atggaatatc aggtagtaat tcctctaagt cataatttcc gtatattctt ttattttttc
     7981 gttttgcttg gtaaagcatt atggttaaat ctgaatttaa ttccttctga ggaatgtatc
     8041 cttgttcata aagctcttgt aaccattctc cataaataaa ttcttgtttg ggaggatgat
     8101 tccacggtac catttcttgc tgaataataa ttgttaattc aatatatcgt aagttgcttt
     8161 tatctcctat tttttttgaa ataggtctaa ttttttgtat aagtatttct ttactttgat
     8221 ctgtcaatgg ttcagatacg acgactaaaa agtcaagatc actatttggt tttagtccac
     8281 tctcaactcc tgatccaaac atgtaagtac caataaggtt attttttaaa tgtttccgaa
     8341 gtattttttt cactttatta atttgttcgt atgtattcaa atatatcctc ctcactattt
     8401 tgattagtac ctattttata tccatagttg ttaattaaat aaacttaatt tagtttattt
     8461 atagatttca ttggcttcta aattttttat ctagataata attattttag ttaattttat
     8521 tctagattat atatgatatg atctttcatt tccataaaac taaagtaagt gtaaacctat
     8581 tcattgtttt aaaaatatct cttgccagtc acgttacgtt attag
//
